Editing of eIF(iso)4E.c confers resistance against Turnip mosaic virus in Brassica rapa

Turnip mosaic virus(TuMV)constitutes one of the primary diseases affecting Brassica rapa,severely impacting its production and resulting in crop failures in various regions worldwide.Recent research has demonstrated the significance of plant translation initiation factors,specifically the eIF4E and eIF4G family genes,as essential recessive disease resistance genes.In our study,we conducted evolutionary and gene expression studies,leading us to identify e IF(iso)4E.c as a potential TuMV-resistant gene.Leveraging CRISPR/Cas9 technology,we obtained mutant B.rapa plants with edited eIF(iso)4E.c gene.We confirmed eIF(iso)4E.c confers resistance against TuMV through phenotypic observations and virus content evaluations.Furthermore,we employed ribosome profiling assays on eif(iso)4e.c mutant seedlings to unravel the translation landscape in response to TuMV.Interestingly,we observed a moderate correlation between the fold changes in gene expression at the transcriptional and translational levels(R^(2)=0.729).Comparative analysis of ribosome profiling and RNA-seq data revealed that plant-pathogen interaction,and MAPK signaling pathway-plant pathways were involved in eIF(iso)4E.c-mediated TuMV resistance.Further analysis revealed that sequence features,coding sequence length,and normalized minimal free energy,influenced the translation efficiency of genes.Our study highlights that the loss of e IF(iso)4E.c can result in a highly intricate translation mechanism,acting synergistically with transcription to confer resistance against TuMV.

Genome-wide identification, characterization, and expression analysis of the SWEET gene family in cucumber

SWEETs （sugars will eventually be exported transporters） are a novel class of recently identified sugar transporters that play important roles in diverse physiological processes. However, only a few species of the plant SWEETgene family have been functionally identified. Up till now, there has been no systematic analysis of the SWEETgene family in Cucurbitaceae crops. Here, a genome-wide characterization of this family was conducted in cucumber（Cucumis sativus L.）. A total of 17 CsSWEETgenes were identified, which are not evenly distributed over the seven cucumber chromosomes. Cucumber SWEET protein sequences possess seven conserved domains and two putative serine phosphorylation sites. The phylo- genetic tree of the SWEET genes in cucumber, Arabidopsis thaliana, and Oryza sativa was constructed, and all the SWEET genes were divided into four clades. In addition, a number of putative cis-elements were identified in the promoter regions of these CsSWEET genes： nine types involved in phytohormone responses and eight types involved in stress responses. Moreover, the transcript levels of CsSWEETgenes were analyzed in various tissues using quantitative real-time polymerase chain reaction. A majority （70.58%） of the CsSWEET genes were confined to reproductive tissue development. Finally, 18 putative watermelon ClaSWEETgenes and 18 melon CmSWEETgenes were identified that showed a high degree of similarity with CsSWEETgenes. The results from this study provided a basic understanding of the CsSWEETgenes and may also facilitate future research to elucidate the function of SWEET genes in cucumber and other Cucurbitaceae crops.

Effects of fresh-cut and storage on glucosinolates profile using broccoli as a case study

Glucosinolates(GLS) contribute to the unique flavour, nutrition, and plant defence of the Cruciferous vegetables. Understanding the GLS changes through postharvest processing is essential for defined preservation. In this study, four different fresh-cut types, whole flower(W),floret(F), quarterly cut floret(QF) and shredded floret(FS) of broccoli, were stored for 0, 1, 2 and 3 day(s) to explore GLS responses to postharvest treatments. As a result, seven GLS were identified, mainly including glucoraphanin(RAA), neoglucobrassicin(NEO), and glucobrassicin(GBC)and accounting for 52.69%, 20.12% and 14.99% of the total GLS(21.92 ± 0.48) μmol · g ^(-1 )DW, respectively. FS had the sharpest decrease in GLS after three days of storage(6.55 ± 0.37) μmol · g-1DW, while QF had the least(10.16 ± 0.33) μmol · g ^(-1 )DW. All GLS components decreased over storage, except for 4-methoxyglucobrassicin(4 ME) in FS and QF, suggesting its key role in serious wound defence. The results suggested certain postharvest approaches influenced the flavour and nutrition of broccoli.

Efficient generation of targeted point mutations in the Brassica oleracea var.botrytis genome via a modified CRISPR/Cas9 system

In this study,we used the modified CRISPR/Cas9 system to produce targeted point mutations in cauliflower.Acetolactate synthase(ALS)and Centromere-specific histone H3 variant(CENH3)genes were selected as the base-editing targets and hypocotyls of cauliflower were used as explants.For ALS gene,a C-to-T conversion in the Pro182 codon(CCT)can alter the encoded amino acid,likely resulting in herbicide resistance,and a C-to-T mutation in the Leu133 codon(CTT)in the CENH3 gene may produce a haploid inducer.Results indicated that the transformation efficiency was 1.8%–4.5%and the mutation efficiencies for the ALS and CENH3 genes were approximately 22%and 87%,respectively.The ALS mutant cauliflower showed strong herbicide resistance,with possible immediate implications for broadleaf weed control in cauliflower fields.

Evaluation of Maturity and Flavor of Melons Using an Electronic Nose

In this work,an electronic nose was used to evaluate the different cultivars and mature stages of melons,so as to establish a scientific method to accurately distinguish the maturity and varieties of melons. Principal component analysis (PCA) and linear discriminant analysis (LDA ) showed that immature melons could be well distinguished from mature melons using electronic nose. When PCA method was used to analyze,electronic nose could completely classify and identify the maturity of melons. Meanwhile,the electronic nose could distinguish different varieties of melons with high discrimination value. The flavor of samples under cut or no cut conditions would slightly change,leading to the variation of discrimination value among different varieties. The samples with similar flavor under no cut condition could be analyzed through cutting mode. The research built a rapid and accurate method to judge the maturity of melons instead of man sense.

Genetic relationship and pedigree of Chinese watermelon varieties based on diversity of perfect SNPs

Watermelon(Citrullus lanatus)is one of the world’s most important fruit crops,and China produces the most watermelons in the world.Recently,a watermelon variome consisting of 414 key resequenced accessions was reported.However,the genetic relationships and pedigree of Chinese watermelon varieties in the seed market remain unclear.In this study,241 evenly distributed perfect single nucleotide polymorphisms(SNPs)derived from the watermelon variome were selected for variety identification.The diversity of 247 Chinese watermelon varieties was identified based on their SNP genotypes.The 247 watermelon varieties were clustered into five subpopulations:the East Asian ecotype,intermediate ecotype,small fruit with red flesh ecotype,small fruit with yellow flesh ecotype,and American ecotype.We further established the pedigree of four subpopulations,of which JingXinNo.1,ZaoChunHongYu,HuangXiaoYu and XiaoLan,and Sugarlee were the main doner of the East Asian ecotype,small fruit with red flesh ecotype,small fruit with yellow flesh ecotype,and American ecotype,respectively.Thirty-two core SNPs were selected and applied in watermelon variety identification.They were also validated by the Kompetitive allele-specific PCR(KASPar)platform.The present study furthered our understanding of the genetic relationships and pedigree of watermelon varieties in China,and will help to manage the plant variety protection in watermelon.

Comparison of DUS testing and SNP fingerprinting for variety identification in cucumber

Variety identification plays an important role in protecting the intellectual property of varieties,ensuring seed quality,and encouraging breeding innovation.Currently,morphological evaluation in the field,such as distinctness,uniformity,and stability(DUS)testing,and DNA fingerprinting in the laboratory using molecular markers are two dominant methods used for variety identification.Few studies have compared the results of these approaches,and the relationship between the two methods is obscure.In this study,134 dominant cucumber varieties were evaluated using 50 DUS testing traits and genotyped by 40 single nucleotide polymorphisms(SNPs).The 40 SNPs were developed in our previous study and arewell suited for variety identification.In the DUS testing,significant positive or negative correlations among 50 DUS traits were observed,and 20 core traits,including 15 fruit traits,were further selected to increase field inspection efficiency.This suggested that fruit shape plays an important role in variety identification.The ratio of fruit length/diameter was themost important trait,explaining 9.2%of the phenotypic variation.In the DNA fingerprinting test,the 40 SNPs were highly polymorphic and could distinguish all of the 134 cucumber varieties,and 14 core SNPs were selected to improve the identification rate.Interestingly,the population structure analysis of 134 cucumber varieties by phenotypic data in the DUS test was in accordance with the genotypic data from the DNA fingerprinting,indicating that all varieties could be divided into the same four subgroups:European type,North China type,South China type,and hybrids of the North China and South China types.Moreover,linear correlativity of distinguishment for each pair of varieties was observed between the DUS test and the DNA fingerprinting.These results indicated that these two methods have good application in future research,especially for the scaled-up analysis of hundreds of varieties.

Inhibitory effect of chitosan on growth of the fungal phytopathogen,Sclerotinia sclerotiorum,and sclerotinia rot of carrot

The antifungal activity of chitosan on a common fungal phytopathogen, Sclerotinia sclerotiorum, and the control effect on sclerotinia rot of carrot were investigated. Mycelial growth and fungal biomass were strongly inhibited by chitosan. Using propidium iodide stain combined with fluorescent microscopy, the plasma membrane of chitosan-treated S. sclerotiorum mycelia was observed to be markedly damaged. Concomitantly, protein leakage and lipid peroxidation was also found to be significantly higher in chitosan-treated mycelia compared to the control. Chitosan provided an effective control of sclerotinia rot of carrot, with induction of activity of defense-related enzymes including polyphenoloxidase and peroxidase. These data suggest that the effects of chitosan on sclerotinia rot of carrot may be associated with the direct damage to the plasma membrane and lipid peroxidation of S. sclerotiorum, and the elicitation of defense response in carrot.

Combined genomic, transcriptomic, and metabolomic an alyses provide in sights into chayote (Sechium edule) evolution and fruit development

Chayote(Sechium edule)is an agricultural crop in the Cucurbitaceae family that is rich in bioactive components.To enhance genetic research on chayote,we used Nanopore third-generation sequencing combined with Hi-C data to assemble a draft chayote genome.A chromosome-level assembly anchored on 14 chromosomes(N50 contig and scaffold sizes of 8.40 and 46.56 Mb,respectively)estimated the genome size as 606.42 Mb,which is large for the Cucurbitaceae,with 65.94%(401.08 Mb)ofthe genome comprising repetitive sequences;28,237 protein-coding genes were predicted.Comparative genome analysis indicated that chayote and snake gourd diverged from sponge gourd and that a whole-genome duplication(WGD)event occurred in chayote at 25±4 Mya.Transcriptional and metabolic analysis revealed genes involved in fruit texture,pigment,fl avor,fl avonoids,antioxidants,and plant hormones during chayote fruit development.The analysis of the genome,transcriptome,and metabolome provides insights into chayote evolution and lays the groundwork for future research on fruit and tuber development and genetic improvements in chayote.

Transcriptomics and metabolomics analyses provide insights into postharvest ripening and senescence of tomato fruit under low temperature

Tomato is one of the most important vegetable crops in the world and is a model plant used to study the ripening of climacteric fleshy fruit.During the ripening process of tomato fruit,flavor and aroma metabolites,color,texture and plant hormones undergo significant changes.However,low temperatures delayed the ripening process of tomato fruit,inhibiting flavor compounds and ethylene production.Metabolomics and transcriptomics analyses of tomato fruit stored under low temperature(LT,5°C)and room temperature(RT,25°C)were carried out to investigate the effects of storage temperature on the physiological changes in tomato fruit after harvest.The results of transcriptomics changes revealed that the differentially expressed genes(DEGs)involved in tomato fruit ripening,including several kinds of transcription factors(TFs)(TCP,WRKY,MYB and bZIP),enzymes involved in cell wall metabolism[beta-galactosidase(β-GAL),pectinesterase(PE)and pectate lyase(PL),cellulose and cellulose synthase(CESA)],enzymes associated with fruit flavor and aroma[acetyltransferase(AT),malic enzyme(ME),lipoxygenase(LOX),aldehyde dehydrogenase(ALDH),alcohol dehydrogenase(ADH)and hexokinase(HK)],genes associated with heat stress protein 70 and genes involved in the production of plant hormones such as Ethylene responsive factor 1(ERF1),Auxin/indoleacetic acids protein(AUX/IAA),gibberellin regulated protein.Based on the above results,we constructed a regulatory network model of the effects of different temperatures during the fruit ripening process.According to the analysis of the metabolomics results,it was found that the contents of many metabolites in tomato fruit were greatly affected by storage temperature,including,organic acids(L-tartaric acid,a-hydroxyisobutyric acid and 4-acetamidobutyric acid),sugars(melezitose,beta-Dlactose,D-sedoheptulose 7-phosphate,2-deoxyribose 1-phosphate and raffinose)and phenols(coniferin,curcumin and feruloylputrescine).This study revealed the effects of storage temperature on postharvest tomato fruit and provided a basis for further understanding of the molecular biology and biochemistry of fruit ripening.

Identification of long noncoding RNAs involved in resistance to downy mildew in Chinese cabbage

Brassica downy mildew,a severe disease caused by Hyaloperonospora brassicae,can cause enormous economic losses in Chinese cabbage(Brassica rapa L.ssp.pekinensis)production.Although some research has been reported recently concerning the underlying resistance to this disease,no studies have identified or characterized long noncoding RNAs involved in this defense response.In this study,using high-throughput RNA sequencing,we analyzed the disease-responding mRNAs and long noncoding RNAs in two resistant lines(T12–19 and 12–85)and one susceptible line(91–112).Clustering and Gene Ontology analysis of differentially expressed genes(DEGs)showed that more DEGs were involved in the defense response in the two resistant lines than in the susceptible line.Different expression patterns and proposed functions of differentially expressed long noncoding RNAs among T12–19,12–85,and 91–112 indicated that each has a distinct disease response mechanism.There were significantly more cis-and trans-functional long noncoding RNAs in the resistant lines than in the susceptible line,and the genes regulated by these RNAs mostly participated in the disease defense response.Furthermore,we identified a candidate resistance-related long noncoding RNA,MSTRG.19915,which is a long noncoding natural antisense transcript of a MAPK gene,BrMAPK15.Via an agroinfiltration-mediated transient overexpression system and virus-induced gene silencing technology,BrMAPK15 was indicated to have a greater ability to defend against pathogens.MSTRG.19915-silenced seedlings showed enhanced resistance to downy mildew,probably because of the upregulated expression of BrMAPK15.This research identified and characterized long noncoding RNAs involved in resistance to downy mildew,laying a foundation for future in-depth studies of disease resistance mechanisms in Chinese cabbage.

Application of droplet digital PCR in detection of seed-transmitted pathogen Acidovorax citrulli

Bacterial fruit blotch caused by Acidovorax citrulli is a serious threat to cucurbit industry worldwide.The pathogen is seedtransmitted,so seed detection to prevent distribution of contaminated seed is crucial in disease management.In this study,we adapted a quantitative real-time PCR(qPCR)assay to droplet digital PCR(ddPCR)format for A.citrulli detection by optimizing reaction conditions.The performance of ddPCR in detecting A.citrulli pure culture,DNA,infested watermelon/melon seed and commercial seed samples were compared with multiplex PCR,qPCR,and dilution plating method.The lowest concentrations detected(LCD)by ddPCR reached up to 2 fg DNA,and 102 CFU mL–1 bacterial cells,which were ten times more sensitive than those of the qPCR.When testing artificially infested watermelon and melon seed,0.1%infestation level was detectable using ddPCR and dilution plating method.The 26 positive samples were identified in 201 commercial seed samples through ddPCR,which was the highest positive number among all the methods.High detection sensitivity achieved by ddPCR demonstrated a promising technique for improving seed-transmitted pathogen detection threshold in the future.

MiR396 is involved in plant response to vernalization and flower development in Agrostis stolonifera

MicroRNA396(miR396)has been demonstrated to regulate flower development by targeting growth-regulating factors(GRFs)in annual species.However,its role in perennial grasses and its potential involvement in flowering time control remain unexplored.Here we report that overexpression of miR396 in a perennial species,creeping bentgrass(Agrostis stolonifera L.),alters flower development.Most significantly,transgenic(TG)plants bypass the vernalization requirement for flowering.Gene expression analysis reveals that miR396 is induced by long-day(LD)photoperiod and vernalization.Further study identifies VRN1,VRN2,and VRN3 homologs whose expression patterns in wild-type(WT)plants are similar to those observed in wheat and barley during transition from short-day(SD)to LD,and SD to cold conditions.However,compared to WT controls,TG plants overexpressing miR396 exhibit significantly enhanced VRN1 and VRN3 expression,but repressed VRN2 expression under SD to LD conditions without vernalization,which might be associated with modified expression of methyltransferase genes.Collectively,our results unveil a potentially novel mechanism by which miR396 suppresses the vernalization requirement for flowering which might be related to the epigenetic regulation of VRN genes and provide important new insight into critical roles of a miRNA in regulating vernalization-mediated transition from vegetative to reproductive growth in monocots.

Genome-wide analysis of changes in miRNA and target gene expression reveals key roles in heterosis for chinese cabbage biomass

Heterosis is a complex phenomenon in which hybrids show better phenotypic characteristics than their parents do.Chinese cabbage(Brassica rapa L.spp.pekinensis)is a popular leafy crop species,hybrids of which are widely used in commercial production;however,the molecular basis of heterosis for biomass of Chinese cabbage is poorly understood.We characterized heterosis in a Chinese cabbage hybrid cultivar and its parental lines from the seedling stage to the heading stage;marked heterosis of leaf weight and biomass yield were observed.Small RNA sequencing revealed 63 and 50 differentially expressed microRNAs(DEMs)at the seedling and early-heading stages,respectively.The expression levels ofthe majority of miRNA clusters in the hybrid were lower than the mid-parent values(MPVs).Using degradome sequencing,we identi fied 1,819 miRNA target genes.Gene ontology(GO)analyses demonstrated that the target genes ofthe MPV-DEMs and low parental expression level dominance(ELD)miRNAs were signi ficantly enriched in leaf morphogenesis,leaf development,and leaf shaping.Transcriptome analysis revealed that the expression levels of photosynthesis and chlorophyll synthesis-related MPV-DEGs(differentially expressed genes)were signi ficantly different in the F_(1) hybrid compared to the parental lines,resulting in increased photosynthesis capacity and chlorophyll content in the former.Furthermore,expression of genes known to regulate leaf development was also observed at the seedling stage.Arabidopsis plants overexpressing BrGRF4.2 and bra-miR396 presented increased and decreased leaf sizes,respectively.These results provide new insight into the regulation of target genes and miRNA expression patterns in leaf size and heterosis for biomass of B.rapa.

Molecularly tagged genes and quantitative trait loci in cucumber with recommendations for QTL nomenclature

Cucumber,Cucumis sativus L.(2n=2x=14),is an important vegetable crop worldwide.It was the first specialty crop with a publicly available draft genome.Its relatively small,diploid genome,short life cycle,and selfcompatible mating system offers advantages for genetic studies.In recent years,significant progress has been made in molecular mapping,and identification of genes and QTL responsible for key phenotypic traits,but a systematic review of the work is lacking.Here,we conducted an extensive literature review on mutants,genes and QTL that have been molecularly mapped or characterized in cucumber.We documented 81 simply inherited trait genes or major-effect QTL that have been cloned or fine mapped.For each gene,detailed information was compiled including chromosome locations,allelic variants and associated polymorphisms,predicted functions,and diagnostic markers that could be used for marker-assisted selection in cucumber breeding.We also documented 322 QTL for 42 quantitative traits,including 109 for disease resistances against seven pathogens.By alignment of these QTL on the latest version of cucumber draft genomes,consensus QTL across multiple studies were inferred,which provided insights into heritable correlations among different traits.Through collaborative efforts among public and private cucumber researchers,we identified 130 quantitative traits and developed a set of recommendations for QTL nomenclature in cucumber.This is the first attempt to systematically summarize,analyze and inventory cucumber mutants,cloned or mapped genes and QTL,which should be a useful resource for the cucurbit research community.

Telomere-to-telomere carrot (Daucus carota) genome assembly reveals carotenoid characteristics

Carrot(Daucus carota)is an Apiaceae plant with multi-colored fleshy roots that provides a model system for carotenoid research.In this study,we assembled a 430.40 Mb high-quality gapless genome to the telomere-to-telomere(T2T)level of“Kurodagosun”carrot.In total,36268 genes were identified and 34961 of them were functionally annotated.The proportion of repeat sequences in the genome was 55.3%,mainly long terminal repeats.Depending on the coverage of the repeats,14 telomeres and 9 centromeric regions on the chromosomes were predicted.A phylogenetic analysis showed that carrots evolved early in the family Apiaceae.Based on the T2T genome,we reconstructed the carotenoid metabolic pathway and identified the structural genes that regulate carotenoid biosynthesis.Among the 65 genes that were screened,9 were newly identified.Additionally,some gene sequences overlapped with transposons,suggesting replication and functional differentiation of carotenoid-related genes during carrot evolution.Given that some gene copies were barely expressed during development,they might be functionally redundant.Comparison of 24 cytochrome P450 genes associated with carotenoid biosynthesis revealed the tandem or proximal duplication resulting in expansion of CYP gene family.These results provided molecular information for carrot carotenoid accumulation and contributed to a new genetic resource.

Effects of a Composite Soil Amendment on the Quality of Pak Choi(Brassica rapa L. Chinensis Group) Growing in Pb-contaminated Soils

In the present study, a pot experiment was carded out to investigate the effects of a composite soil amendment at different levels （0, 600, 900 and 1 200 mg/kg） on the soil organic matter content, pH, Pb uptake and nutritional quality of pak choi （Brassica rapa L. Chinensis Group） grown in the soils contaminated by four levels （800, 1 200, 1 600 and 2 000 mg/kg） of Pb. The results showed that 900 mg/kg soil amendment increased the soil organic matter content by 24.19%- 60.00%, but had no significant influence on soil pH. Comparing with control （without amendment application）, the security and nutritional quality of pak choi were obviously improved by applying the soil amendment. In detail, 900 mg/kg soil amendment improved the content of Vc of pak choi growing in 800, 1 200, 1 600 and 2 000 mg/kg Pb-contaminated soils by 13.27%-69.30%, the content of soluble sugar by 54.17%- 87.50%, the dry matter weight by 28.36%-33.39%, and decreased the content of Pb by 19.11%-35.72% and the content of crude fiber by 20.83%-31.03%, respectively. These data indicated that the composite amendment can be used for the in situ repair of Pb-contaminated soils, and the recommended dosage is 900 mg/kg.

The genome and transcriptome analysis of snake gourd provide insights into its evolution and fruit development and ripening

Snake gourd(Trichosanthes anguina L.),which belongs to the Cucurbitaceae family,is a popular ornamental and food crop species with medicinal value and is grown in many parts of the world.Although progress has been made in its genetic improvement,the organization,composition,and evolution of the snake gourd genome remain largely unknown.Here,we report a high-quality genome assembly for snake gourd,comprising 202 contigs,with a total size of 919.8 Mb and an N50 size of 20.1 Mb.These findings indicate that snake gourd has one of the largest genomes of Cucurbitaceae species sequenced to date.The snake gourd genome assembly harbors 22,874 protein-coding genes and 80.0%of the genome consists of repetitive sequences.Phylogenetic analysis reveals that snake gourd is closely related to sponge gourd but diverged from their common ancestor~33–47 million years ago.The genome sequence reported here serves as a valuable resource for snake gourd genetic research and comparative genomic studies in Cucurbitaceae and other plant species.In addition,fruit transcriptome analysis reveals the candidate genes related to quality traits during snake gourd fruit development and provides a basis for future research on snake gourd fruit development and ripening at the transcript level.

Analysis on Composition and Content of Glucosinolates in Broccoli Flowers and Leaves

Glucosinolate composition and content were evaluated in flowers and leaves of 12 different broccoli varieties. The results indicated that there were 9 glucosinolates in broccoli,namely Glucoiberin(IBE); Progoitrin(PRO);Sinigrin(SIN);Glucoraphanin(RAA);Gluconapin(NAP);4-Hydroxyglucobrassicin(4OH);Glucobrassicin(GBC);4-Methoxyglucobrassicin(4ME);Neoglucobrassicin(NEO).Total glucosinolate content in flowers was 1-5 times higher than in leaves. The predominant glucosinolate in broccoli was glucoraphanin.

Glucosinolate Variation in Turnip (Brassica rapa L.) at Different Growing Stages

The seed, leaf and root parts of three turnip （ Brassica rapa L.） cultivars were analyzed for glucosinolates. The component and concentration of glucosinolates in the three cultivars were also analyzed at different growing stages. Eight kinds of glucosinolates were tested in turnip. They were PRO, NAP, 4OH, GBN, GBC, NAS, 4ME and NEO, respectively. The contents and kinds of glucosinolates varied at different growing stages. The content of NAS was the highest in the seed, GBC and NEO contents were the highest in the seedling, GBN and NEO contents were the highest in the leaf, while the highest NAS content was in the root.